Publications by authors named "Junwen Ma"

14 Publications

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Global transcriptomic analysis of functional oligosaccharide metabolism in Pediococcus pentosaceus.

Appl Microbiol Biotechnol 2021 Feb 29;105(4):1601-1614. Epub 2021 Jan 29.

Key Laboratory of Food Bioengineering (China National Light Industry), College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, China.

Lactic acid bacteria (LAB) are important in food fermentation and may enhance overall host health. Previous studies to explore LAB metabolism mainly focused on the genera Lacticaseibacillus and Lactococcus. Pediococcus pentosaceus, historically recognized as an important food fermentation bacterial strain, can produce bacteriocins and occasionally demonstrated probiotic functionalities. This study thoroughly surveyed the growth kinetic of three P. pentosaceus isolates in various culture formulations, especially in fructooligosaccharide (FOS), xylooligosaccharide (XOS), or konjac mannooligosaccharide (KMOS) conditions. Results showed that P. pentosaceus effectively metabolized KMOS, the culture of which led to 23.6-fold population increase. However, FOS and XOS were less metabolized by P. pentosaceus. On functional oligosaccharide cultures, P. pentosaceus could result in higher population proliferation, more acidified fermentation environment, and higher glycoside hydrolysis activities in the culture. RNA-Seq analysis classified 1572 out of 1708 putative genes as mRNA-coding genes. The dataset also revealed that the three functional oligosaccharides led to extensive global functional gene regulations. Phosphate conservation and utilization efficiency enhancement may serve as a leading transcriptional regulation direction in functional oligosaccharide metabolisms. In summary, these discovered metabolic characteristics could be employed to support future studies. KEY POINTS: • Konjac mannooligosaccharides effectively promoted P. pentosaceus proliferation. • Functional genes were highly regulated in functional oligosaccharide utilization. • Phosphate conservation was an important transcriptional regulation direction.
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http://dx.doi.org/10.1007/s00253-021-11120-5DOI Listing
February 2021

Ginkgoic acid impedes gastric cancer cell proliferation, migration and EMT through inhibiting the SUMOylation of IGF-1R.

Chem Biol Interact 2021 Mar 25;337:109394. Epub 2021 Jan 25.

Department of General Surgery, Xi'an Gaoxin Hospital, Xi'an, 710075, China. Electronic address:

The imbalance of SUMOylation is related to different cancers, including gastric cancer (GC). Ginkgolic acid (GA) inhibits the growth and invasion of many cancer cells, and it has been reported to restrain SUMOylation. However, the role of GA in GC and whether it functions through SUMOylation remains to be clarified. Our research revealed that GA (15:1) inhibited cell proliferation, migration, epithelial-mesenchymal transition (EMT) and overall protein SUMOylation in BGC823 and HGC27 cells. In addition, knockdown of SUMO1 (small ubiquitin-like modifier) instead of SUMO2/3 played a similar role to GA in cell behaviors. Besides, nuclear IGF-1R (insulin-like growth factor 1 receptor) expression was markedly upregulated in GC cells compared to normal gastric epithelial cells. GA prevented IGF-1R from binding to SUMO1, thereby suppressing its nuclear accumulation. Further research found that IGF-1R directly bound to SNAI2 (snail family zinc finger 2) promoter. The interference of IGF-1R downregulated the mRNA and protein levels of SNAI2, while the overexpression of SUMO1, IGF-1R and UBC9 (SUMO-conjugating enzyme) played the opposite role. Furthermore, the co-transfection of SUMO1, UBC9 and IGF-1R vectors or the overexpression of SNAI2 reversed the inhibitory effects of GA on cell proliferation, migration and EMT. Finally, GA impeded the growth of GC xenografts and decreased the expression of nuclear IGF-1R and SNAI2 in vivo. In conclusion, these findings demonstrated that GA hindered the progression of GC by inhibiting the SUMOylation of IGF-1R. Thus, GA might be a promising therapeutic for GC.
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http://dx.doi.org/10.1016/j.cbi.2021.109394DOI Listing
March 2021

Biochemical Properties of a Novel D-Mannose Isomerase from Pseudomonas syringae for D-Mannose Production.

Appl Biochem Biotechnol 2021 Jan 23. Epub 2021 Jan 23.

Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Engineering, China Agricultural University, No.17 Qinghua Donglu, Haidian District, Beijing, 100083, China.

D-Mannose isomerase can reversibly catalyze D-fructose to D-mannose which has various beneficial effects. A novel D-mannose isomerase gene (PsMIaseA) from Pseudomonas syringae was cloned and expressed in Escherichia coli. The recombinant D-mannose isomerase (PsMIaseA) showed the highest amino acid sequence homogeneity of 50% with ManI from Thermobifda fusca. PsMIaseA was purified through Ni-NTA chromatography, and its specific activity was 818.6 U mg. The optimal pH and temperature of PsMIaseA were pH 7.5 and 45 °C, respectively. The enzyme was stable within a wide pH range from 5.0 to 10.0. It could efficiently convert D-fructose to D-mannose without any metal ions. When PsMIaseA was incubated with 600 g/L D-fructose for 6 h, the space-time yield of D-mannose reached 27.2 g L h with a maximum conversion ratio of 27%. Therefore, the D-mannose isomerase may be suitable for green production of D-mannose.
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http://dx.doi.org/10.1007/s12010-021-03487-yDOI Listing
January 2021

Occurrence and distribution of five antibiotic resistance genes during the loading period in sludge treatment wetlands.

J Environ Manage 2020 Nov 6;274:111190. Epub 2020 Aug 6.

College of Environment and Resources, Dalian Minzu University, Dalian, 116600, China.

The objectives of this study were to clarify the distribution as well as the removal mechanism of antibiotic resistance genes (ARGs) within three sludge treatment wetlands (STWs) during a loading period of two years. Three STW units were constructed and run during the loading period: Unit 1 (U1) built with aeration tubes, Unit 2 (U2) built with aeration tubes and reeds, and Unit 3 (U3) built with reeds only. All targeted ARGs, intI1, and 16S rRNA were detected in residual sludge in the order of magnitude: 16S rRNA>sul1>intI1>sul2>tetC>tetA>ermB. The abundance of the five targeted ARGs, intI1, and 16S rRNA increased in residual sludge, during the loading period, which may be due to the increase in bacteria caused by the continuous import of exogenous nutrients. However, STWs can also remove ARGs from sewage during the loading period and the mean removal efficiency of five resistance genes was 73.0%. The removal rates of intI1 and 16S rRNA were 73.5% and 78.6%, respectively. Positive correlations were detected in abundance of most ARGs and intI1, as well as 16S rRNA (P < 0.05), indicating intI1 plays a vital part in the propagation of ARGs. The removal of bacteria harboring these genes also occurs in the STW units.
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http://dx.doi.org/10.1016/j.jenvman.2020.111190DOI Listing
November 2020

LINC02163 promotes colorectal cancer progression via miR-511-3p/AKT3 axis.

Artif Cells Nanomed Biotechnol 2020 Dec;48(1):961-968

Department of Gastroenterology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.

Long non-coding RNAs and microRNAs are functional regulators in tumour progression. Herein, we revealed the level LINC02163 was up-regulated in CRC tissues and cell lines, and the expression of LINC02163 negatively correlated with prognosis of CRC patients. Functional experiments demonstrated knockdown of LINC02163 significantly attenuated CRC cells proliferation and metastasis. Mechanism analysis showed miR-511-3p could bind LINC02163 and , and the expressional level of miR-511-3p negatively correlated with the abundance of LINC02163 and . Inhibition of LINC02163 suppressed cell proliferation, while transfection of miR-511-3p inhibitor or AKT3 in LINC02163-depletion cells restored cell growth and abolished the cell cycle arrest in G0/G1 phase. Therefore, it was indicated that LINC02163 exerted pro-tumour effect through miR-511-3p/AKT3 axis and was prognostic marker for colorectal cancer.
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http://dx.doi.org/10.1080/21691401.2020.1773486DOI Listing
December 2020

Biochemical characterization of a novel α-L-fucosidase from Pedobacter sp. and its application in synthesis of 3'-fucosyllactose and 2'-fucosyllactose.

Appl Microbiol Biotechnol 2020 Jul 9;104(13):5813-5826. Epub 2020 May 9.

Key Laboratory of Food Bioengineering (China National Light Industry), College of Food Science and Nutritional Engineering, China Agricultural University, No.17 Qinghua Donglu, Haidian District, Beijing, 100083, China.

Fucosyllactoses have gained much attention owing to their multiple functions, including prebiotic, immune, gut, and cognition benefits. In this study, human milk oligosaccharide (HMO) 2'-fucosyllactose (α-L-Fuc-(1,2)-D-Galβ-1,4-Glu, 2'FL) and its isomer 3'-fucosyllactose (α-L-Fuc-(1,3)-D-Galβ-1,4-Glu, 3'FL) with potential prebiotic effect were synthesized efficiently by a novel recombinant α-L-fucosidase. An α-L-fucosidase gene (PbFuc) from Pedobacter sp. CAU209 was successfully cloned and expressed in Escherichia coli (E. coli). The deduced amino acid sequence shared the highest identity of 36.8% with the amino sequences of other reported α-L-fucosidases. The purified α-L-fucosidase (PbFuc) had a molecular mass of 50 kDa. The enzyme exhibited specific activity (26.3 U/mg) towards 4-nitrophenyl-α-L-fucopyranoside (pNP-FUC), 3'FL (8.9 U/mg), and 2'FL (3.4 U/mg). It showed the highest activity at pH 5.0 and 35 °C, respectively. PbFuc catalyzed the synthesis of 3'FL and 2'FL through a transglycosylation reaction using pNP-FUC as donor and lactose as acceptor, and total conversion ratio was up to 85% at the optimized reaction conditions. The synthesized mixture of 2'FL and 3'FL promoted the growth of Lactobacillus delbrueckii subsp. bulgaricus NRRL B-548, L. casei subsp. casei NRRL B-1922, L. casei subsp. casei AS 1.2435, and Bifidobacterium longum NRRL B-41409. However, the growths of E. coli ATCC 11775, S. enterica AS 1.1552, L. monocytogenes CICC 21635, and S. aureus AS 1.1861 were not stimulated by the mixture of 2'FL and 3'FL. Overall, our findings suggest that PbFuc possesses a great potential for the specific synthesis of fucosylated compounds.Key Points• A novel α-L-fucosidase (PbFuc) from Pedobacter sp. was cloned and expressed.• PbFuc showed the highest hydrolysis activity at pH 5.0 and 35 °C, respectively.• It was used for synthesis of 3'-fucosyllactose (3'FL) and 2'-fucosyllactose (2'FL).• The mixture of 3'FL and 2'FL promoted the growth of some Lactobacillus sp. and Bifidobacteria sp.
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http://dx.doi.org/10.1007/s00253-020-10630-yDOI Listing
July 2020

Biochemical Characterization and Structural Analysis of a β--Acetylglucosaminidase from for Efficient Production of -Acetyl-d-glucosamine.

J Agric Food Chem 2020 May 11;68(20):5648-5657. Epub 2020 May 11.

College of Food Science and Nutritional Engineering, China Agricultural University, No.17 Qinghua East Road, Haidian District, Beijing 100083, China.

Bioproduction of -acetyl-d-glucosamine (GlcNAc) from chitin, the second most abundant natural renewable polymer on earth, is of great value in which chitinolytic enzymes play key roles. In this study, a novel glycoside hydrolase family-18 β--acetylglucosaminidase (PbNag39) from suitable for GlcNAc production was identified and biochemically characterized. It possessed a unique shallow catalytic groove (5.8 Å) as well as a smaller C-terminal domain (solvent-accessible surface area, 5.1 × 10 Å) and exhibited strict substrate specificity toward -acetyl chitooligosaccharides (COS) with GlcNAc as the sole product, showing a typical manner of action of β--acetylglucosaminidases. Thus, an environmentally friendly bioprocess for GlcNAc production from ball-milled powdery chitin by an enzyme cocktail reaction was further developed. By using the new route, the powdery chitin conversion rate increased from 23.3% (v/v) to 75.3% with a final GlcNAc content of 22.6 mg mL. The efficient and environmentally friendly bioprocess may have great application potential in GlcNAc production.
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http://dx.doi.org/10.1021/acs.jafc.9b08085DOI Listing
May 2020

Biochemical characterization of a bifunctional chitinase/lysozyme from Streptomyces sampsonii suitable for N-acetyl chitobiose production.

Biotechnol Lett 2020 Aug 13;42(8):1489-1499. Epub 2020 Mar 13.

Key Laboratory of Food Bioengineering (China National Light Industry), College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083, China.

Objectives: Chitinases play important role in chitin bioconversion, while few of them have been put into use due to their poor properties. We aimed to identify and characterize chitinases suitable for N-acetyl chitooligosaccharides (COSs) production from chitin materials.

Results: A chitinase gene (SsChi28) from Streptomyces sampsonii XY2-7 was cloned and heterologously expressed in E. coli BL21 (DE3) as an active protein. The deduced protein shared high sequence identities and structure similarities with some glycoside hydrolase family 19 chitinases. The recombinant enzyme (SsChi28) was purified and biochemically characterized. SsChi28 was a monomeric protein with a molecular mass of 30 kDa estimated by SDS-PAGE. It was most active at pH 6.0 and 55 °C, respectively, and stable in a wide pH range of 3.5-11.5 and up to 60 °C. The enzyme exhibited strict substrate specificities towards ethylene glycol chitin (222.3 U/mg) and colloidal chitin (20.1 U/mg). Besides, it displayed lysozyme activity against Micrococcus lysodeikticus. SsChi28 hydrolyzed colloidal chitin to yield mainly N-acetyl chitobiose, accounting high up to 73% (w/w) in total products.

Conclusion: The excellent enzymatic properties of SsChi28 may make it potential in chitin bioconversion (especially for N-acetyl COS production), as well as in biological control of fungal diseases.
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http://dx.doi.org/10.1007/s10529-020-02834-zDOI Listing
August 2020

Evaluation of the fate of nutrients, antibiotics, and antibiotic resistance genes in sludge treatment wetlands.

Sci Total Environ 2020 Apr 30;712:136370. Epub 2019 Dec 30.

School of Environment Science & Technology, Dalian University of Technology, Dalian 116024, China.

The aim of this research was to analyze the elimination of nutrients, antibiotics as well as antibiotic resistance genes (ARGs) in different sludge treatment wetlands (STWs) with or without reeds and aeration tubes. Five antibiotics, including oxytetracycline, tetracycline, azithromycin, sulfamethoxazole, and sulfadiazine; five ARGs, including two tetracycline ARGs (tetC and tetA), one macrolide ARGs (ermB), and two sulfonamide ARGs (sul1 and sul2); and one integrase gene (intI1) were determined in the surface and bottom layers of three STWs, respectively. The removal efficiencies of antibiotics in the bottom layer were lower than that in the surface layer, while the elimination efficiencies of ARGs showed opposite trend. Strong correlations were observed among the contents of antibiotics as well as related ARGs, and the abundance of ARGs had a strong correlation with intI1. The results demonstrated that the contents of these pollutants decreased during the resting period in all the STWs, while the wetland had reeds and aeration tubes performed the best.
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http://dx.doi.org/10.1016/j.scitotenv.2019.136370DOI Listing
April 2020

Curdlan () (1→3)-β-d-Glucan Oligosaccharides Drive M1 Phenotype Polarization in Murine Bone Marrow-Derived Macrophages via Activation of MAPKs and NF-κB Pathways.

Molecules 2019 Nov 22;24(23). Epub 2019 Nov 22.

Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.

Functional oligosaccharides, particularly curdlan (1→3)-β-d-glucan oligosaccharides (GOS), play important roles in modulating host immune responses. However, the molecular mechanisms underlying the immunostimulatory effects of GOS on macrophage polarization are not clear. In this work, GOS (5-1000 µg/mL) were non-toxic to bone marrow-derived macrophages (BMDMs) with improved pinocytic and bactericidal capacities. Incubation with GOS (100 µg/mL) induced M1 phenotype polarization of BMDMs as evidenced by increased CD11c/CD86 (10.1%) and M1 gene expression of inducible nitric oxide synthase, interleukin (IL)-1β, and chemokine C-C-motif ligand 2. Accordingly, the secretion of cytokines IL-1β, IL-6, monocyte chemotactic protein-1, and tumor necrosis factor-α, as well as the nitrite release of BMDMs were increased by GOS (100 µg/mL). Expression of mitogen-activated protein kinases (MAPKs) of phosphorylated (p)-c-Jun amino-terminal kinase, p-extracellular signal regulated kinase, and p-p38 in BMDMs were increased by GOS, as well as the p-Stat1. Moreover, nuclear factor-kappa B (NF-κB) p-p65 expression in BMDMs was promoted by GOS while it suppressed IκBα expression. Receptor blocking with anti-CR3 (CD11b/CD18) and anti-toll-like receptor (TLR) 2 antibodies diminished GOS induced M1 phenotype polarization with reduced mRNA expression of M1 genes, decreased cytokine and nitrite releases, and suppressed signaling pathway activation. Thus, CR3 (CD11b/CD18) and TLR2 mediated activation of MAPKs and NF-κB pathways are responsible for GOS induced polarization of BMDMs.
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http://dx.doi.org/10.3390/molecules24234251DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6930549PMC
November 2019

Fate of antibiotics in three distinct sludge treatment wetlands under different operating conditions.

Sci Total Environ 2019 Jun 11;671:443-451. Epub 2019 Mar 11.

School of Environment Science & Technology, Dalian University of Technology, Dalian 116024, China.

Sludge treatment wetlands (STWs) have recently been used to treat surplus sludge. However, the distribution of antibiotics involved in the process has not been comprehensively investigated. This study aimed to evaluate the fate of two antibiotics, i.e., ciprofloxacin (CIP) and azithromycin (AZM) in STWs during the treatment of surplus sludge. Three pilot-scale STWs units-S1 with aeration tubes, S2 with aeration tubes and reed planting, and S3 with reed planting-were constructed and operated under feeding followed by resting periods. The results showed that antibiotic content in residual sludge decreased over time and unit S2 performed the best in terms of antibiotic removal. Planting reed considerably improved the antibiotic removal performance of the STWs. Biodegradation and absorption resulted in removal of most of the antibiotics in the test units. Less than 2% of the antibiotics was taken up by plants, whereas <5% of the influent antibiotics left the STW units through the drainage discharge. Overall, STW units contributed to effectively decrease CIP and AZM to 41-72% and 49-84%, respectively.
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http://dx.doi.org/10.1016/j.scitotenv.2019.03.147DOI Listing
June 2019

Fate of antibiotics and the related antibiotic resistance genes during sludge stabilization in sludge treatment wetlands.

Chemosphere 2019 Jun 25;224:502-508. Epub 2019 Feb 25.

School of Environment Science & Technology, Dalian University of Technology, Dalian, 116024, China.

Antibiotics contamination and related antibiotics resistance genes (ARGs) in wastewater sludge have been a concern for environmental pollution and human health risk for years. This study investigated the fate of antibiotics and related ARGs in sludge from sludge treatment wetlands (STWs). We examined three sludge treatment beds i.e. unit No.1 was sludge drying bed with aeration tubes; unit No.2 was a ventilated sludge drying reed bed; and the unit No.3 was a sludge drying reed bed without aeration tubes. The targeted antibiotics included oxytetracycline, roxithromycin and azithromycin. The targeted ARGs included tetA, tetC, msrSA and ermB. The results indicated that in all three units antibiotics were removed significantly and related ARGs declined over one year period. The antibiotics concentrations in the surface layer were lower than those in the bottom layer. The highest removal efficiency of the targeted antibiotics was observed in the unit No.2. The removal efficiency of the targeted ARGs influenced by different parameters, especially reeds, aeration tubes and temperature. Correlation analysis between the concentrations of antibiotics and corresponding ARGs showed that in both the bottom and surface layer of all three units, a significant correlation (p < 0.05) was found between the concentrations of roxithromycin and azithromycin and the absolute abundances of msrSA. A significant correlation (p < 0.05) was also observed between the concentrations of oxytetracycline and the absolute abundances of tetA and tetC. The results demonstrated that STWs can effectively reduce the target antibiotics contents and related ARGs; and the STW with reed and aeration tubes performed better.
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http://dx.doi.org/10.1016/j.chemosphere.2019.02.168DOI Listing
June 2019

Fractalkine/CX3CR1 axis modulated the development of pancreatic ductal adenocarcinoma via JAK/STAT signaling pathway.

Biochem Biophys Res Commun 2017 12 3;493(4):1510-1517. Epub 2017 Oct 3.

Department of Hepatobiliary Surgery, General Hospital of NingXia Medical University, Yinchuan, NingXia, 750004, China.

Pancreatic ductal adenocarcinoma (PDAC) is a fatal malignancy with an estimated 5 year survival rate of approximately 5% of all stages combined. High potential of PDAC metastasis is a leading cause for high mortality and poor prognosis. The majority of patients present with distant metastasis at diagnosis. Fractalkine (FKN) is recognized as a chemokine and a specific ligand of CX3CR1. It has been reported that FKN/CX3CR1 system was upregulated in many types of solid tumors. However, role of FKN/CX3CR1 in PDAC development remains unclear. In the current investigation, we found that FKN and CX3CR1 expression was significantly increased in PDAC tissues, especially in the metastatic samples, and was highly-correlated with severity of PDAC. Ectopic expression of FKN promoted the proliferation and migration of PDAC, while knockdown of CX3CR1 reversed the function of FKN. In addition, PDAC cells with FKN-deficiency showed impaired proliferation and migration activity. The underlying mechanism is that FKN/CX3CR1 activated JAK/STAT signaling, which in turn regulated cell growth. Consistently, in vivo tumorigenesis assay validated the regulatory role of FKN/CX3CR1 in PDAC growth. Our investigation helped understanding the pathogenesis of PDAC occurrence, and demonstrated critical role of FKN/CX3CR1 in PDAC development.
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http://dx.doi.org/10.1016/j.bbrc.2017.10.006DOI Listing
December 2017

siRNA-based targeting of fractalkine overexpression suppresses inflammation development in a severe acute pancreatitis rat model.

Int J Mol Med 2012 Sep 29;30(3):514-20. Epub 2012 Jun 29.

Department of Gastroenterology, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, P.R. China.

Fractalkine (FKN), a chemokine that acts as both an adhesion molecule and a chemoattractant, is expressed in many inflammatory diseases. Chemokines play a crucial role in severe acute pancreatitis (SAP). This study used adenovirus-mediated siRNA to target FKN overexpression and assessed its ability to suppress inflammation development in a SAP rat model. Adenovirus-mediated FKN siRNA was transfected into cerulein-stimulated AR42J cells. The growth of cerulein-stimulated AR42J cells was determined by colony formation and MTT assays. The inhibitory effect of the FKN siRNA was studied in a SAP rat model in vivo and detected by ELISA, RT-PCR, western blot analysis and immunohistochemistry. FKN, IL-8 and TNF-α were found to be overexpressed in cerulein-stimulated AR42J cells by ELISA and western blot analysis (P<0.05). The animal experiments confirmed that FKN siRNA could inhibit inflammation development in SAP. The values of serum FKN, TNF-α and IL-8 levels were decreased after FKN siRNA treatment (P<0.05). Furthermore, western blotting and RT-PCR analysis showed that FKN protein and mRNA levels were decreased after injection with FKN siRNA (P<0.05). Immunohistochemistry also showed that inflammation was decreased after injection with FKN-siRNA in the SAP rat model. Treatment with siRNA can inhibit FKN overexpression and also suppresses inflammation development in a SAP rat model. More importantly, this study indicated that FKN, which is overexpressed in the SAP rat model, may serve as a novel and effective therapeutic target for SAP.
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http://dx.doi.org/10.3892/ijmm.2012.1050DOI Listing
September 2012